The ACOD1 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from human NCI-H1299 non-small cell lung carcinoma (NSCLC) cells. These cells feature targeted disruption of the ACOD1 gene, which encodes the itaconate-producing enzyme cis-aconitate decarboxylase. This polyclonal pool provides a heterogeneous loss-of-function model, minimizing clonal selection bias. The product is tailored for research into metabolism-inflammation interplay in cancer.
NCI-H1299 is an epithelial cell line established from a lymph node metastasis of human NSCLC. It harbors a homozygous partial deletion of TP53, resulting in p53-null status that promotes tumorigenicity and genomic instability. The cells retain lung epithelial characteristics and are highly amenable to transfection and lentiviral transduction, making them a robust host for CRISPR/Cas9 gene editing. This background is widely used to study oncogenic signaling, apoptosis resistance, and metabolic reprogramming in lung cancer.
ACOD1 (immune-responsive gene 1) catalyzes the decarboxylation of cis-aconitate to itaconate, requiring divalent metal ions such as Mg2? or Mn2?. Itaconate acts as a potent metabolic regulator: it inhibits succinate dehydrogenase (SDH), leading to succinate accumulation and TCA cycle modulation, and alkylates KEAP1 to activate NRF2-driven antioxidant responses. ACOD1 transcription is induced by TNF-??, LPS, IFN-??, and type I interferons via NF-??B, IRF1, and STAT1. Downstream, itaconate suppresses the NLRP3 inflammasome, restrains NF-??B signaling, and influences ATF3, I??B??, and glycolytic enzymes GAPDH and LDHA. Thus, ACOD1 integrates immunometabolic control.
Disruption of ACOD1 in NCI-H1299 cells eliminates itaconate production, releasing SDH inhibition and potentially amplifying succinate-driven inflammatory pathways. In a p53-null background, loss of itaconate may intensify NF-??B and NLRP3 inflammasome activity, alter cytokine output, and shift metabolism toward glycolysis. This knockout model enables detailed examination of how ACOD1-derived itaconate shapes tumor-intrinsic inflammatory circuits and paracrine effects within the tumor microenvironment, facilitating investigation of immunometabolite roles in lung cancer progression.
Applications include itaconate quantification via LC-MS, ACOD1 expression analysis by western blotting and RT-qPCR, and metabolic profiling with Seahorse assays. The cells suit NF-??B luciferase reporter studies and cytokine arrays to probe inflammatory signaling, as well as invasion and migration assays for metastatic behavior. RNA-seq can uncover transcriptional changes upon ACOD1 loss. The model also supports drug screening for itaconate pathway modulators and co-culture studies of tumor-immune interactions. For further information or to request a quote, contact Ascent Research.
